Method of treating rhinitis or sinusitis

ABSTRACT

The present invention is directed to methods of treating mucosal inflammation associated with rhinitis or sinusitis by administering peptidases that recognize and cleave polypeptides at Xaa-Pro sequences. In addition, the invention encompasses therapeutic packages in which pharmaceutical compositions containing the peptidases are preloaded in a device suitable for intranasally delivering drug.

FIELD OF THE INVENTION

[0001] The present invention is directed to methods of treating inflammation associated with rhinitis or sinusitis by intranasally delivering peptidases to a patient. The invention also encompasses therapeutic packages in which a peptidase is preloaded in a device designed for intranasally delivering drug.

BACKGROUND OF THE INVENTION

[0002] Rhinitis, an inflammation of the nasal mucosal membrane, is characterized by sneezing, rhinorrhea, nasal congestion, and increased nasal secretion. It is often accompanied by sinusitis, an inflammation of the sinuses. When these conditions persist for a period of more than three weeks, they are termed “chronic.” More than 37 million Americans, particularly those with allergies or asthma, suffer from these conditions, making them the most common chronic medical problems in the United States.

[0003] Chronic “rhinosinusitis” or sinusitis is difficult to treat successfully. In general, treatment consists of a combination of antibiotics and decongestants or antihistamines. In addition, steroid nasal sprays are commonly used to reduce inflammation. For patients with severe chronic sinusitis, oral steroids, such as prednisone, may also be prescribed. However, the long-term safety of steroid administration, especially in children, is not fully understood and oral steroids often have significant side effects. When drug therapy fails, surgery is usually the only alternative.

[0004] The mucosal tissue lining the nasal and sinus passages is densely packed with sensory neurons (Alving, et al., Cell Tissue Res. 264:529-538 (1991); Saria, et al., Am. Rev. Respir. Dis. 147:1330-1335 (1988)). When activated, these neurons release a variety of bioactive peptides that contribute to inflammation by causing vasodilation, stimulating mucosal gland secretion, and promoting infiltration by inflammatory mast cells, lymphocytes and eosinophils (Stead, et al., Immunol. Rev. 10:333-359 (1987); Mygind, et al., Eur. J. Respir. Dis. 64(S128):1-379 (1983)). Included among the released bioactive peptides are substance P, calcitonin-gene related peptide, neuropeptide Y and vasoactive intestinal peptide (Lundblad, et al., Acta. Physiol. Scand. 529:1-42 (1984)). Means for controlling the activity of these peptides should provide an effective treatment for the inflammation associated with both rhinitis and sinusitis.

SUMMARY OF THE INVENTION

[0005] The present invention stems from the discovery that dipeptidyl peptidase IV, an exopeptidase that cleaves Xaa-Pro dipeptides from the N-terminus of polypeptides, is present in human nasal mucosa at levels that are inversely related to inflammation. Thus, low levels of dipeptidyl peptidase IV are associated with a high density of inflammatory cells, and high levels of dipeptidyl peptidase IV are associated with a low density of inflammatory cells. This is important because dipeptidyl peptidase IV degrades peptides that contribute to the pathophysiology of rhinitis, sinusitis and asthma (Mentlein, et al., Regul. Peptides 49:133-144 (1993); Heymann, et al., FEBS Lett. 91:360-364 (1978); Beauvais, et al., Fum. Infect. Immun. 65:3042-3047 (1997)). Among the inflammation-related peptides cleaved are NPY, SP, and desArg1 bradykinin. Based upon these observation and further experiments, the concept has been developed that the intranasal administration of dipeptidyl peptidase IV can reduce inflammation in the mucosal tissue that lines both the nasal cavity and sinuses. Other proteases that possess the same proteolytic activity should also produce a positive therapeutic effect. These proteases include quiescent cell proline dipeptidase (Underwood, et al., J. Biol. Chem. 274:34053-34058 (1999)), dipeptidyl peptidase 8 (Abbott, et al., Eur. J. Biochem. 20 267:6140-6150 (2000)), and attractin (Duke-Cohan, et al., Proc. Nat'l. Acad. Sci. U.S.A. 95:11336-11341 (1998)).

[0006] In its first aspect, the invention is directed to a method of treating a patient for inflammation of the nasal or sinus mucosa. The method involves intranasally administering a therapeutically effective amount of a peptidase (preferably an exopeptidase) that cleaves at Xaa-Pro residues, where Xaa represents any of the 20 amino acids commonly found in animals. A “therapeutically effective” dose is defined as an amount sufficient to produce a significant reduction in inflammation as evidence by either a reduced number of inflammatory cells in mucosal tissue or by a significant improvement in one or more symptoms associated with the inflammation. For example, in the case of rhinitis or sinusitis, a therapeutically effective dose would be a sufficient amount to produce a significant reduction in sneezing, coughing, sinus-related headaches, nasal obstruction, mucosal secretion, or other discomfort associated with these conditions. Inflammatory cells include mast cells, lymphocytes and eosinophils. In general, it is expected that a therapeutically effective dose for any of the proteases used will be between I microgram and 1 milligram and, typically, between 5 micrograms and 500 micrograms.

[0007] The preferred peptidase for use in the method is dipeptidyl peptidase IV. Other peptidases that can be used include quiescent cell proline dipeptidase; dipeptidyl peptidase 8, and attractin. In each case, it is the human form of the peptidase that is preferred. However, peptidases from other species (e.g., that secreted by Aspergilus Fumigatus, see Examples section) may also be used provided that they have the ability to cleave at the Xaa-Pro sequence. Although the method will work for rhinitis and sinusitis caused by any disease or condition, it is expected that the most common causes will be allergies or asthma.

[0008] In another aspect, the invention is directed to a therapeutic package in which a device for intranasally delivering drug to a patient is preloaded with a solution or powder containing one or more of the peptidases described above. The invention is compatible with any intranasal delivery device (including encapsulated dosage forms) and with any of the numerous compositions that have been described for delivering drugs by means of the nasal cavity. When liquid compositions are used in the device, it is expected that peptidase will be present at a concentration of between 1 μg/ml and 10 mg/ml, and more typically, at a concentration of between 10 μg/ml. and 1 mg/ml.

[0009] The invention also encompasses the concept that SP is particularly important in causing inflammation in lung and nasal mucosa. Any method that reduces the local activity of this peptide should be useful in the treatment of rhinitis or sinusitis. A reduction in activity may be accomplished either using a peptidase that degrades SP (e.g., one of the peptidases described above) or by administering an agent that inhibits the binding of SP to the NK1 receptor (see Examples section).

DETAILED DESCRIPTION OF THE INVENTION

[0010] A. Preparation of Peptidases

[0011] The present invention is directed to treatment methods which utilize peptidases that have the common characteristic of cleaving at Xaa-Pro sites. These may be purchased commercially or obtained using any of the procedures described in the relevant literature. For example, the gene corresponding to the peptidase can be isolated and used for recombinant protein production. Especially preferred peptidases, along with references relevant to their isolation and recombinant production, are: human dipeptidyl peptidase IV, shown herein as SEQ ID NO: 1 (Misumi, et al., Biochim. Biophys. Acta 15:1131 (1992); Darmoul, et al., J Biol. Chem. 267:4824-4833 (1992); Abbott, et al., Immunogenetics 40:331-338 (1994)); human quiescent cell proline dipeptidase, shown herein as SEQ ID NO: 2 (Underwood, et al., J. Biol. Chem. 274:34053-34058 (1999)); human attractin, shown herein as SEQ ID NO: 3 (Duke-Cohan, et al., Proc. Nat'l. Acad. Sci. USA 95:11336-11341 (1998); Nagase, et al., DNA Res. 5:31-39 (1998)); and human dipeptidyl peptidase 8, shown herein as SEQ ID NO: 4 (Abbott, et al., Eur. J Biochem. 267:6140-6150 (2000)). In addition to being made recombinantly, these proteins can be synthesized using methods that are well-known in the art.

[0012] B. Making of Pharmaceutical Compositions

[0013] Compositions for intranasally delivering peptidases can be made in accordance with methods that are standard in the art (see, e.g., Remington's Pharmaceutical Sciences, 16^(th) ed. A. Oslo Editor, Easton Pa. (1980)). Enzymes will typically be prepared in admixture with conventional excipients. Suitable carriers may include, but are not limited to: water; salt solutions; alcohols; vegetable oils; polyethylene glycols; gellatin; carbohydrates such as lactose, amylose or starch; talc; hydroxymethylcellulose etc. The pharmaceutical preparations can be sterilized and, if desired, mixed with auxiliary agents such as preservatives or stabilizers. The invention is compatible with any of the numerous compositions that have been disclosed in the art for nasal delivery including those in: U.S. Pat. Nos. 6,054,462; 4,946,870; 5,897,858; and 4,476,116. The concentration of peptidase present can vary over a wide range. Typically, in liquid formulations, peptide should be present in an amount of between 1 μg/ml and 10 mg/ml and, more commonly, at a concentration of between 10 μg/ml and 1 mg/ml.

[0014] Treatment Methods

[0015] The present invention is directed to methods for treating inflammation present in the mucosal lining of the nasal or sinus passages. It depends upon the direct administration of sufficient peptidase to proteolytically cleave peptides known to contribute to inflammation. The total dosage of peptidase to be administered to a patient should be at least the amount required to achieve this objective as reflected by a reduction or elimination of symptoms associated with inflammation. For example, a patient being treated for rhinosinusitis should receive sufficient compound to reduce or eliminate the frequency or intensity of sinus headache, reduce coughing, congestion, sneezing, respiratory obstruction or other discomforts associated with the condition.

[0016] In general, a patient may begin by self-administering a relatively small dose of compound and then repeat administration as necessary. For example, a patient may begin by administering 0.1 mg per day and then increase the dosage upward using changes in inflammation-related symptoms as a guide. Typically, it is expected that patients will receive a daily dose of between 1 μg and I mg per day, and, more typically, between 5 μg and 500 μg. Daily dosages may be provided in either a single or multiple regimen with the latter being generally preferred. These are simply guidelines, since the actual dose will be determined by the patient and their physician based upon a variety of clinical factors.

[0017] Therapeutic Packages

[0018] In addition to the pharmaceutical compositions described above, the invention includes therapeutic packages for the intranasal delivery of the compositions. A therapeutic package is comprised of a device designed for the intranasal inhalation of medication which has been preloaded with a pharmaceutical composition containing one or more of the peptidases described above. In general, spray devices are preferred, such as those disclosed in U.S. Pat. No. 6,145,703; WO 95/00195; U.S. Pat. No. 5,307,953; EP 0388651; U.S. Pat. Nos. 4,017,007; 5,301,846; 3,176,883; or 4,286,735. Devices for administering powders or nasal drops may also be used. When the pharmaceutical composition is in the form of a solution, it is expected that peptidase should generally be present at a concentration of between 1 μg/ml and 10 mg/ml, and, more typically, at a concentration of between 10 μg/ml and 1 mg/ml.

EXAMPLES

[0019] I. Materials and Methods

[0020] Recombinant Dipeptidyl Peptidase IV (DPPIV)

[0021] A soluble form of DPPIV secreted from Aspergilus Fumigatus has been previously characterized and was used in the present experiments (Beauvais et al., Infection Immun. 65 3042-3047 (1997)). The enzyme has an apparent molecular weight of 95 kDa. It was expressed in the yeast Pichia Pastoris and purified to more than 99% purity as assessed by electrophoresis and gel-filtration to reach a specific activity of 40 units/mg of protein. SP (1.8 μg) co-incubated with 0.016 μg of DPPIV for ten minutes at 37° C. is degraded to SP5-11 as identified by mass-spectrometry. If an excess of SP is added (3.5 μg) a partial of digestion of the peptide is observed.

[0022] Patients

[0023] Forty-five patients, 23 males and 22 females, suffering from nasal obstruction, rhinorrhea and headaches for more than eighteen months were included in the study. Pre-operative rhinoscopy revealed septal deviation associated with concha bullosa of the middle turbinate. All patients underwent septoplasty and partial middle turbinectomy under endoscopic control with general anesthesia. The age range was 14 to 64 years with the average patient being 39 years of age. Patients with allergy, nasal polyps or tumors were excluded.

[0024] Tissue Processing

[0025] Samples of middle turbinate mucosa from patients undergoing partial turbinectomy were fixed immediately in ice cold acetone with 2 mM phenyl methyl sulphonyl fluoride and 20 mM iodoacetamide and incubated overnight at −20° C. Biopsies were embedded in glycol methacrylate resin and allowed to polymerize overnight at 4° C.

[0026] Antibodies

[0027] The following monoclonal antibodies were used: CD26 (clone BA5, DAKO) directed against DPPIV protease, diluted 1:20; CD1A (Biogenex) for dendritic cells, diluted 1:20; CD31 (DAKO) recognizing the adhesion molecule PECAM on endothelial cells, diluted 1:20; and the polyclonal antibody CD3 (DAKO) directed against T cells and used at a dilution of 1:20.

[0028] Immunohistochemical Staining

[0029] Serial sections, 2 mm thick, were cut using a Reichert-ung microtome equipped with a glass knife. Immunohistochemical staining was performed using the streptavidin biotin-peroxidase method with aminoethyl-carbazole (AEC) as substrate.

[0030] Quantification of Inflammation in Nasal Biopsies and Intensity of Symptoms

[0031] Mucosal samples of the middle turbinate from both sides (N=90) were fixed in formaldehyde and dehydrated, embedded in paraffin, and colored by haematoxylin-eosin. They were then examined under a Zeiss microscope at 40× magnification. Histological analysis included defining the integrity of the pseudo-stratified columnar epithelium, noting the presence or absence of edema and quantifying the number of inflammatory cells within the submucosa. This was accomplished using a scale graded from 0 to +++, where 0 meant no inflammatory cells and +++ represented abundant inflammatory cells. Using the Rank Spearman correlation test, the correlation between DPPIV and the degree of inflammation of the nasal mucosa was examined. The intensity of nasal obstruction, rhinorrhea and headache was recorded by means of a visual analog scale graded from 0 to 5, where 0 corresponds to the absence of symptoms and 5 corresponds to severely intense symptoms. Nasal airway resistance was recorded by means of anterior rhinomanometry (rhinotest).

[0032] The Determination of DPPIV Activity in Human Mucosa Biopsies

[0033] DPPIV activity was determined according to Scharpe, et al. (Clin. Chim. Acta 195: 125-132 (1990)) with the following modifications: nasal biopsies were sonicated in the presence of 0.5 ml of 100 mM Tris-HCl, pH 8, for 2 minutes on ice using a Branson sonifier (output 4) and centrifuged for 10 minutes at 15,000 rpm in a microfuge at 4° C. The supernatant was recovered and the pellet was treated with 0.5 ml of 100 mM Tris-HCl, pH8, containing 2% Triton X100. The suspension was vortexed for one minute and centrifuged for 10 minutes at 15,000 rpm in a microfuge at 4° C. The supernatant was recovered, pooled with the one obtained previously, and stored at −20° C. DPPIV activity was determined on 1, 2.5 and 5 μl of supernatant fluorometrically using Gly-Pro-AMC (Novabiochem) at 5 mM final concentration for 60 minutes at 37° C. under agitation in an Eppendorf thermomixer in 25 μl of 100 mM Tris-HCl, pH 8. The reaction was stopped by the addition of 2.5 μl of pure acetic acid. The incubation mixture was recovered in 3 ml of water. A blank value was obtained by incubating the substrate in the absence of enzyme and a standard curve was determined using AMC fluorescence measurement on a fluorometer. The DPPIV activities were standardized based on wet tissue weight and specific activities expressed as pmoles of substrate converted per mg of tissue per minute.

[0034] Experiments in the Pig In Vivo

[0035] Experiments were performed on pigs of both sexes (body weight 18-25 kg). All animals were premedicated with atropine (0.05 mg/kg) and ketamine (20 mg/kg i.m.). They were anesthesized with thiopentone (5 mg/kg i.v.). After tracheostomy, animals were intubated and artificially ventilated by a volume regulated ventilator. During surgery, animals were given a continuous i.v. infusion of Ringer's solution, pancuronium bromide (0.25 mg/kg) and heparin. Each experiment lasted approximately 8 hours. Catheters were placed in the femoral artery for systemic blood pressure and heart rate monitoring and in the femoral vein for thiopentone, heparin and fluid administration (300 ml/hr). The contralateral femoral vein was canulated for blood sampling. Surgical preparation of the internal maxillary artery was done in accordance with Lacroix (Acta Physiol. Scand. 136:1-63 (1989)). Selective recording of the nasal arterial blood flow was performed with a Transonic flow probe (probe 2.4RB 143) of 2.4 mm diameter placed around the sphenopalatine artery. The flow probe was connected to a T202S ultrasonic blood flowmeter. All the arterial branches situated downstream of the flow probe were ligated and cut except for the superficial temporal artery which was cannulated with a PE 90 catheter for infusions or injections.

[0036] Variations in the area under the curve (AUC) of the sphenopalatine artery vascular resistance (Vr), derived from both mean arterial blood pressure and systemic blood flow curves, were analyzed over time. Durations of the responses of the vascular resistance were compared and all results were expressed in percent of baseline. Vascular parameters were recorded simultaneously using a 6 pen trace recorder.

[0037] All animals were intranasally administered DPIV (50 μg, 26.5 pmoles/kg), subjected to sympathetic nerve stimulation (SNS, 15V, 5 ms, 10 Hz for 2 minutes) and then infused with histamine (0.1-25 μg), capsacin (0.01-25 μg), bradykinin (0.001-10 μg), SP or its C-terminal fragment SP 5-11, DPPIV, or NK1 antagonist L733060 in the superficial temporal artery under controlled conditions. In each case, the vascular response of the animal was measured. This entire procedure was then repeated after local i.a. pretreatment with the alpha-adrenergic receptor antagonist phenoxybenzanime (1 mg/kg).

[0038] Vascular Responses to DPPIV in Domestic Pigs In Vivo

[0039] The basal blood flow in the internal maxillary artery of the pig under control conditions was 4.92±0.7 ml min⁻¹ kg⁻¹. After section of the sympathetic nerves on the left side, the homolateral nasal arterial flow was 6.1±0.26 ml min⁻¹ kg⁻¹ (representing a 19.34%±4.3% increase). The mean arterial blood pressure (MAP) was not modified by the section of the sympathetic nerves. Electrical stimulation of the sympathetic nerve induced a frequency-depended increase of the SVR whereas the MAP was not significantly modified. Sympathetic nerve stimulation at 10 Hz for 5 minutes reduced the blood flow in the maxillary artery from 4.92±0.7 ml min⁻¹ kg⁻¹ to 1.40±0.47 ml min⁻¹ kg⁻¹, representing an increase in SVR of 71.5% and lasting more than 6 minutes. The i.a. infusion of phenoxybenzamine induced a MAP reduction of 10±2%. The basal blood flow in the sphenopalatine artery, the heart rate and the MAP were not significantly affected by the administration of exogenous dipeptidyl peptidase IV.

[0040] Statistical Analysis

[0041] All values were expressed in terms of mean ±SEM. Statistical analysis was done by analysis of variance, ANOVA. A value of p<0.05 was taken as statistically significant.

[0042] II. Results

[0043] DPPIV is Expressed in Endothelial Cells and Submucosal Seromucus Glands from Patients

[0044] Using immunohistochemistry, DPPIV-like immunoreactivity (-LI) was detected in submucosal seromucus glands and leukocytes. In submucosal seromucus glands, DPPIV-LI appeared to be located in the apical position. Endothelial cells in blood vessels expressed weak DPPIV-LI. Some epithelial cells located in the human nasal mucosa were also DPPIV positive. However, these cells did not show any positive immunoreactivity for CD1A and Protein 100, suggesting they were not Langerhans cells.

[0045] Correlation Between DPPIV and Inflammation in the Human Nasal Mucosa

[0046] Nasal mucosa biopsies were sampled from both nostrils in 45 patients suffering from rhinosinusitis to determine if DPPIV activity was affected by the mucosal inflammation. DPPIV activity was found to vary from undetectable to 707 pmol/min/mg. Histological analysis revealed marked differences in the density of inflammatory cells within the submucosa of the nasal biopsies studied. A low activity of DPPIV was associated with a high density of inflammatory cells in the nasal mucosa of patients with chronic rhinosinusitis. When the density of inflammatory cells observed was plotted against DPPIV activity, the regressive correlation was found to be statistically significant (p<0.001). Nasal mucosa samples obtained from smokers exhibited a significantly lower DPPIV activity than samples obtained from non-smokers having the same low density of inflammatory cells (p<0.01).

[0047] When the density of inflammatory cells in each biopsy was plotted against the subjective evaluation of nasal obstruction, the data fitted a statistically significant correlation (p<0.01), suggesting that nasal inflammation increased in parallel with subjective nasal obstruction severity. In addition, nasal mucosa biopsies were obtained from 10 patients with significant improvement of their symptoms (p<0.001), 6 months after endonasal surgery. The DPPIV activity was significantly increased in all of the samples studied when compared to the preoperative state (p<0.001) indicating that DPPIV activity can be restored when chronic rhinosinusitis is cured. Since there was a clear negative correlation between DPPIV activity and nasal mucosal inflammation, the hemodynamic effect of SP in the presence of recombinant DPPIV was studied in pig nasal mucosa.

[0048] DPPIV Modulates Inflammatory Response Mediated by Histamine in Pigs

[0049] Histamine is responsible for the early-phase allergic reaction and exerts both direct and indirect effects on sensory nerves, glands and blood vessels of the nasal mucosa (Alving, Acta. Physiol. Scand. 597:1-64 (1991)). There is direct evidence that histamine can release peptides from capsaicin sensitive sensory nerves in the lung by activation of H_(I)-receptors (Alving, et al., Acta. Physiol. Scand. 138:49-60 (1990)). In addition, endogenous or exogenous histamine stimulates sensory fibers, possibly by acting on specific receptors to increase the release of CGRP and SP (Tani, et al., Neurosci. Lett. 115:171-176 (1990).

[0050] Histamine i.a. local injection following sympathetic nerve stimulation (SNS) at 10 Hz for 5 minutes showed a significant reduction in duration at 5.4×10⁻⁰⁹ moles (33%) and at 1.4×10⁻⁰⁷ moles (32%) when compared to control conditions. After i.a. administration of 50 μg of DPPIV and SNS, histamine challenge showed no significant variation in the area under the curve (AUC) or vascular resistance (Vr) when compared to control. However, the duration of histamine effect was reduced by 33.7% at a dose of 5.4×10⁻⁰⁸ moles (p<0.05) and by 50.7% at a dose of 1.4×10⁻⁰⁷ moles (p<0.01). After phenoxybenzamine (PBZ) pretreatment, DPPIV and SNS, histamine vasodilatory effects were markedly reduced. When compared to control, the AUC showed a reduction of 81.9% at a dose of 5.4×10⁻⁰⁸ moles (p<0.001) and a reduction of 81.8% at a dose of 1.4×10⁻⁰⁷ moles (p<0.001). Vr showed a reduction of 67.9% at 5.4×10⁻⁰⁸ moles (p<0.01) and a reduction of 61.1% at 1.4×10⁻⁰⁷ moles (p<0.05). Duration of effect was also reduced at doses of 5.4×10⁻⁰⁹ moles (42.6%, p<0.01); 5.4×10⁻⁰⁸ moles (55.6%, p<0.001); and 1.4×10⁻⁰⁷ moles (62.5%, p<0.001).

[0051] DPPIV Modulates the Vasodilation Evoked by Capsaicin in Pigs

[0052] Capsaicin (8-methyl-N-vanillyl-6-nonenamide), by activating unmylinated sensory C-fibers, acts in many respects like histamine. Capsaicin has been shown to release tachykinins (Hua, et al., Neurosci. 19:313-319 (1986)) as well as CGRP (Stjarne, et al., Regul. Pept. 33:251-262 (1991)) both in vitro and in vivo. Capsaicin injection following SNS showed a dose-dependent decrease of Vr similar that to observed under control conditions. The vasodilatory effect of capsaicin after SNS and PBZ pretreatment showed no significant variation at any doses. However, the duration of vasodilation evoked at 8.2×10⁻⁰⁸ moles was significantly reduced (39.9%, p<0.01) and AUC was reduced by 26.6% at 8.2×10⁻⁰⁸ moles (p<0.05). Capsaicin effects were also modified after DPPIV and SNS. The AUC observed at a dose of 8.2×10⁻⁰⁸ moles showed a reduction by 56.6% (p<0.01). The Vr response was reduced by 34.8% (p<0.05) and the duration of effect at doses of 3.3×10⁻⁰⁹ moles and 8.2×10⁻⁰⁸ moles was reduced by 55.4% (p<0.05) and 50.5% (p<0.001) respectively. After PBZ, DPPIV and SNS, there was marked reduction in Vr, duration of effect, and AUC. Vr at 8.2×10⁻⁰⁸ moles was reduced by 39.9% (p<0.01). At 3.3×10⁻⁰⁹ moles and 8.2×10⁻⁰⁸ moles, the duration of the vascular response was reduced by 64.3% (p<0.05) and 56.8% (p<0.001). At 8.2×10⁻⁰⁸ moles, AUC was reduced by 66.7% (p<0.01).

[0053] DPPIV Modulates the Vasodilatory Response Mediated by Bradykinin in Pigs

[0054] Bradykinin (BK) is a polypeptide involved in nociception and humoral regulation of vascular tonicity and permeability. BK produces marked vasodilation, increases capillary permeability and is involved in most inflammatory reactions, including rhinitis (Svensson, et al., J. Allergy Clin. Immunol. 85:828-833 (1990)). Like histamine and capsaicin, BK stimulates nociceptive sensory nerves to produce CGRP and SP (Hua, et al., Can. J. Physiol., Pharmacol. 73:999-1006 (1995); Vasco, et al., J. Neurosci. 14:4987-4997 (1994)).

[0055] BK injection following SNS showed a dose-dependent decrease of Vr similar to that observed under control conditions. After DPPIV and SNS, BK showed only variation in the duration of effect at doses of 9.4×10⁻¹¹ moles (34%, p<0.01) and 9.4×10⁻⁰⁹ moles (38.3%, p<0.001). After PBV, DPPIV and SNS, vascular parameters were reduced to a smaller extent than that seen with histamine and capsaicin. For example, AUC after i.a. administration of BK was reduced by 29.3% at 9.4×10⁻⁰⁹ moles (p<0.01) and by 40% at 9.4×10⁻¹¹ moles (p<0.01). At 9.4×10⁻¹⁰ moles duration of effect was reduced by 32.2% (p<0.05) and at 9.4×10⁻⁰⁹ moles, duration was reduced by 39.4% (p<0.001).

[0056] DDPIV Modulates the Vasodilatory Response Mediated by SP in Pigs

[0057] Repeated injection of high doses of SP caused a reproducible decrease in sphenopalatine vascular resistance, indicating that the desensitization of neurokinin (NK) receptors did not occur. Therefore, the same animal was used to perform dose-response curves with SP both before and after pretreatment with DPPIV. A dose-response curve for duration of vasodilation and area under the curve was constructed using doses of SP in the range of 1 pg to 0.1 μg. A reproducible dose-response curve was obtained over a 45 minute period using 6 doses of SP. Vasodilation ranged from 15.1% ±3.3% to 37.3% ±3.3% after local intraarterial administration of 1 pg and 100 ng of SP respectively. To determine whether vasodilation evoked by SP was affected by pretreatment with DPPIV, two doses of SP (100 pg and 1 ng) were administered and a reduction in SVR of 13.1% and 14.9% was observed. The same animals received 50 μg (530 pmoles) of DPPIV and similar doses of SP were injected. Recombinant DPPIV had no vascular effect per se. Subsequent administration of the SP resulted in a dramatic diminution of SP-evoked vasodilatory response (by 66 and 71% respectively) when compared to controls. Since DPPIV cleaves SP into SP 5-11, experiments were conducted to determine whether SP 5-11 is also capable of decreasing SVR. Similar to what was observed with respect to SP, a dose response curve was obtained with 10 μg to 1 μg of SP 5-11 (5 to 35% decrease in SVR respectively). However, the vasodilatory effect of SP 5-11 was considerably lower when compared to SP. Based on molarity, SP is 200 fold more potent than SP 5-11 as a vasodilator. Therefore, DPPIV administration to pigs might result in an almost immediate conversion of SP 5-11.

[0058] To further characterize the nature of neurokinin receptor subtype involved in the reduction of SVR, pigs were pretreated with a NK1 antagonist, L-733060, at 114 nmoles/kg to evaluate the blockade of SP-induced decreased in SVR. It was found that the antagonist has a vasoconstrictive effect. A clear inhibition of the SP effect was observed at all doses of SP used except 100 ng. Thus, the SP effect on SVR is likely to mediated by the NK1 receptor.

[0059] III. Discussion

[0060] The localization of the enzyme DPPIV in the apical position of nasal exocrine cells in seromucus glands suggests a role of this enzyme in the protective function of nasal mucus. The presence of DPPIV in vascular endothelial cells and T-cells is in line with earlier histochemical studies of DPPIV in mammalian tissue (Sannes, J. Histochem. Cytochem. 31:684-690 (1983)) as well as with more recent reports on the distribution of human DPPIV (Van. Der. Elden, et al., Clin. Exp. Allergy 28:110-120 (1998)). DPPIV immunoreactivity was also observed in some intraepithelium cells of nasal mucosa. These cells were probably not Langerhans cells since they did not express CD1A or Protein 100 immunoreactivity on their surface.

[0061] Correlation between DPPIV activity and inflammation of nasal mucosa showed a marked decrease of enzyme activity in the presence of severe inflammation. In agreement with Van der Velden, et al. (Id.), who showed that DPPIV activity is reduced in the serum of healthy smokers compared to non-smokers, DPPIV activity was reduced in nasal samples with severe inflammation. In contrast, high DPPIV enzymatic activity was correlated with a small number of inflammatory cells in the nasal mucosa. The fact that enzyme activity increased after treatment of chronic rhinosinusitis is also consistent with the involvement of the enzyme in this pathology. Pretreatment with 50 ug of DPPIV significantly decreased duration of vasodilation and spenopalatine artery vascular resistance when DPPIV was used alone or with the adrenoreceptor blocker phenoxybenzamine.

[0062] DPPIV significantly decreased both peak and duration of the vasodilation evoked by SP. It was found that the SP 5-11 was at least 100 fold less potent than SP at causing vasodilation. These results suggest that a lost of DPPIV expression occurs during chronic rhinosinusitis and that the resulting reduction of SP degradation contributes to the maintenance of nasal mucosa inflammation. In this regard, Nieber, et al. have shown that allergic patients have higher baseline levels of SP-ir in nasal lavage fluids than non-allergic controls and that intranasal allergen increases SP levels in nasal lavage fluids only in subjects with grass pollen allergy (J. Allergy Clin. Immunol. 4 pt1:646-652 (1992)). The antibody used for the SP assay in the Nieber study required both an intact N- and C-terminus peptide sequence. The data is in line with that presented herein and could be explained by the fact that allergic patients expressing less DDPIV than normal volunteers produce intact SP which could be measured by their antibody. In contrast, normal volunteers cleave their SP into SP 5-11 fragments which are no longer detected in the SP assay. SP effects on the vascular bed of the nasal mucosa are maintained only when allergenic stimulation persists in subjects with nasal allergies (i.e., subjects without DPPIV).

[0063] In parallel to SP cleavage by DPPIV into SP 5-11, a N-terminal fragment, SP 1-4, is generated and has been reported to exert an antagonist affect on SP action (Sakurada, et al., J. Pharm. Sci. 88: 1127-1132 (1999)). Thus, DPPIV might modulate SP action, not only by producing a less active NK1 agonist, but also by interfering with SP effects. Since SP 5-11 (but not SP) is subject to cellular uptake, DPPIV may also act to terminate the action of SP via conversion to a form which is removed by cellular processes (Nakata, et al., J. Neurochem. 37:1529-1534 (1981)). Once SP is cleaved by DPPIV into SP 5-11, this fragment is further processed with a high efficiency by aminopeptidase M into inactive fragments. Thus, DPPIV first converts SP to a much less active form, SP 5-11, which is then completely inactivated by the action of aminopeptidase M. Finally, DPPIV could potentially act upon other peptides which are known to be involved in promoting inflammation and which have structures that lend themselves to degradation by this enzyme.

[0064] All references cited are fully incorporated by reference. Having now fully described the invention, it will be understood by those of skill in the art that the invention may be performed within a wide and equivalent range of conditions, parameters and the like, without affecting the spirit or scope of the invention or any embodiment thereof.

1 4 1 766 PRT Homo sapiens 1 Met Lys Thr Pro Trp Lys Val Leu Leu Gly Leu Leu Gly Ala Ala Ala 1 5 10 15 Leu Val Thr Ile Ile Thr Val Pro Val Val Leu Leu Asn Lys Gly Thr 20 25 30 Asp Asp Ala Thr Ala Asp Ser Arg Lys Thr Tyr Thr Leu Thr Asp Tyr 35 40 45 Leu Lys Asn Thr Tyr Arg Leu Lys Leu Tyr Ser Leu Arg Trp Ile Ser 50 55 60 Asp His Glu Tyr Leu Tyr Lys Gln Glu Asn Asn Ile Leu Val Phe Asn 65 70 75 80 Ala Glu Tyr Gly Asn Ser Ser Val Phe Leu Glu Asn Ser Thr Phe Asp 85 90 95 Glu Phe Gly His Ser Ile Asn Asp Tyr Ser Ile Ser Pro Asp Gly Gln 100 105 110 Phe Ile Leu Leu Glu Tyr Asn Tyr Val Lys Gln Trp Arg His Ser Tyr 115 120 125 Thr Ala Ser Tyr Asp Ile Tyr Asp Leu Asn Lys Arg Gln Leu Ile Thr 130 135 140 Glu Glu Arg Ile Pro Asn Asn Thr Gln Trp Val Thr Trp Ser Pro Val 145 150 155 160 Gly His Lys Leu Ala Tyr Val Trp Asn Asn Asp Ile Tyr Val Lys Ile 165 170 175 Glu Pro Asn Leu Pro Ser Tyr Arg Ile Thr Trp Thr Gly Lys Glu Asp 180 185 190 Ile Ile Tyr Asn Gly Ile Thr Asp Trp Val Tyr Glu Glu Glu Val Phe 195 200 205 Ser Ala Tyr Ser Ala Leu Trp Trp Ser Pro Asn Gly Thr Phe Leu Ala 210 215 220 Tyr Ala Gln Phe Asn Asp Thr Glu Val Pro Leu Ile Glu Tyr Ser Phe 225 230 235 240 Tyr Ser Asp Glu Ser Leu Gln Tyr Pro Lys Thr Val Arg Val Pro Tyr 245 250 255 Pro Lys Ala Gly Ala Val Asn Pro Thr Val Lys Phe Phe Val Val Asn 260 265 270 Thr Asp Ser Leu Ser Ser Val Thr Asn Ala Thr Ser Ile Gln Ile Thr 275 280 285 Ala Pro Ala Ser Met Leu Ile Gly Asp His Tyr Leu Cys Asp Val Thr 290 295 300 Trp Ala Thr Gln Glu Arg Ile Ser Leu Gln Trp Leu Arg Arg Ile Gln 305 310 315 320 Asn Tyr Ser Val Met Asp Ile Cys Asp Tyr Asp Glu Ser Ser Gly Arg 325 330 335 Trp Asn Cys Leu Val Ala Arg Gln His Ile Glu Met Ser Thr Thr Gly 340 345 350 Trp Val Gly Arg Phe Arg Pro Ser Glu Pro His Phe Thr Leu Asp Gly 355 360 365 Asn Ser Phe Tyr Lys Ile Ile Ser Asn Glu Glu Gly Tyr Arg His Ile 370 375 380 Cys Tyr Phe Gln Ile Asp Lys Lys Asp Cys Thr Phe Ile Thr Lys Gly 385 390 395 400 Thr Trp Glu Val Ile Gly Ile Glu Ala Leu Thr Ser Asp Tyr Leu Tyr 405 410 415 Tyr Ile Ser Asn Glu Tyr Lys Gly Met Pro Gly Gly Arg Asn Leu Tyr 420 425 430 Lys Ile Gln Leu Ser Asp Tyr Thr Lys Val Thr Cys Leu Ser Cys Glu 435 440 445 Leu Asn Pro Glu Arg Cys Gln Tyr Tyr Ser Val Ser Phe Ser Lys Glu 450 455 460 Ala Lys Tyr Tyr Gln Leu Arg Cys Ser Gly Pro Gly Leu Pro Leu Tyr 465 470 475 480 Thr Leu His Ser Ser Val Asn Asp Lys Gly Leu Arg Val Leu Glu Asp 485 490 495 Asn Ser Ala Leu Asp Lys Met Leu Gln Asn Val Gln Met Pro Ser Lys 500 505 510 Lys Leu Asp Phe Ile Ile Leu Asn Glu Thr Lys Phe Trp Tyr Gln Met 515 520 525 Ile Leu Pro Pro His Phe Asp Lys Ser Lys Lys Tyr Pro Leu Leu Leu 530 535 540 Asp Val Tyr Ala Gly Pro Cys Ser Gln Lys Ala Asp Thr Val Phe Arg 545 550 555 560 Leu Asn Trp Ala Thr Tyr Leu Ala Ser Thr Glu Asn Ile Ile Val Ala 565 570 575 Ser Phe Asp Gly Arg Gly Ser Gly Tyr Gln Gly Asp Lys Ile Met His 580 585 590 Ala Ile Asn Arg Arg Leu Gly Thr Phe Glu Val Glu Asp Gln Ile Glu 595 600 605 Ala Ala Arg Gln Phe Ser Lys Met Gly Phe Val Asp Asn Lys Arg Ile 610 615 620 Ala Ile Trp Gly Trp Ser Tyr Gly Gly Tyr Val Thr Ser Met Val Leu 625 630 635 640 Gly Ser Gly Ser Gly Val Phe Lys Cys Gly Ile Ala Val Ala Pro Val 645 650 655 Ser Arg Trp Glu Tyr Tyr Asp Ser Val Tyr Thr Glu Arg Tyr Met Gly 660 665 670 Leu Pro Thr Pro Glu Asp Asn Leu Asp His Tyr Arg Asn Ser Thr Val 675 680 685 Met Ser Arg Ala Glu Asn Phe Lys Gln Val Glu Tyr Leu Leu Ile His 690 695 700 Gly Thr Ala Asp Asp Asn Val His Phe Gln Gln Ser Ala Gln Ile Ser 705 710 715 720 Lys Ala Leu Val Asp Val Gly Val Asp Phe Gln Ala Met Trp Tyr Thr 725 730 735 Asp Glu Asp His Gly Ile Ala Ser Ser Thr Ala His Gln His Ile Tyr 740 745 750 Thr His Met Ser His Phe Ile Lys Gln Cys Phe Ser Leu Pro 755 760 765 2 492 PRT Homo sapiens 2 Met Gly Ser Ala Pro Trp Ala Pro Val Leu Leu Leu Ala Leu Gly Leu 1 5 10 15 Arg Gly Leu Gln Ala Gly Ala Arg Arg Ala Pro Asp Pro Gly Phe Gln 20 25 30 Glu Arg Phe Phe Gln Gln Arg Leu Asp His Phe Asn Phe Glu Arg Phe 35 40 45 Gly Asn Lys Thr Phe Pro Gln Arg Phe Leu Val Ser Asp Arg Phe Trp 50 55 60 Val Arg Gly Glu Gly Pro Thr Phe Phe Tyr Thr Gly Asn Glu Gly Asp 65 70 75 80 Val Trp Ala Phe Ala Asn Asn Ser Gly Phe Val Ala Glu Leu Ala Ala 85 90 95 Glu Arg Gly Ala Leu Leu Val Phe Ala Glu His Arg Tyr Tyr Gly Lys 100 105 110 Ser Leu Pro Phe Gly Ala Gln Ser Thr Gln Arg Gly His Thr Glu Leu 115 120 125 Leu Thr Val Glu Gln Ala Leu Ala Asp Phe Ala Glu Leu Leu Arg Ala 130 135 140 Leu Arg Arg Asp Leu Gly Ala Gln Asp Ala Pro Ala Ile Ala Phe Gly 145 150 155 160 Gly Ser Tyr Gly Gly Met Leu Ser Ala Tyr Leu Arg Met Lys Tyr Pro 165 170 175 His Leu Val Ala Gly Ala Leu Ala Ala Ser Ala Pro Val Leu Ala Val 180 185 190 Ala Gly Leu Gly Asp Ser Asn Gln Phe Phe Arg Asp Val Thr Ala Asp 195 200 205 Phe Glu Gly Gln Ser Pro Lys Cys Thr Gln Gly Val Arg Glu Ala Phe 210 215 220 Arg Gln Ile Lys Asp Leu Phe Leu Gln Gly Ala Tyr Asp Thr Val Arg 225 230 235 240 Trp Glu Phe Gly Thr Cys Gln Pro Leu Ser Asp Glu Lys Asp Leu Thr 245 250 255 Gln Leu Phe Met Phe Ala Arg Asn Ala Phe Thr Val Leu Ala Met Met 260 265 270 Asp Tyr Pro Tyr Pro Thr Asp Phe Leu Gly Pro Leu Pro Ala Asn Pro 275 280 285 Val Lys Val Gly Cys Asp Arg Leu Leu Ser Glu Ala Gln Arg Ile Thr 290 295 300 Gly Leu Arg Ala Leu Ala Gly Leu Val Tyr Asn Ala Ser Gly Ser Glu 305 310 315 320 His Cys Tyr Asp Ile Tyr Arg Leu Tyr His Ser Cys Ala Asp Pro Thr 325 330 335 Gly Cys Gly Thr Gly Pro Asp Ala Arg Ala Trp Asp Tyr Gln Ala Cys 340 345 350 Thr Glu Ile Asn Leu Thr Phe Ala Ser Asn Asn Val Thr Asp Met Phe 355 360 365 Pro Asp Leu Pro Phe Thr Asp Glu Leu Arg Gln Arg Tyr Cys Leu Asp 370 375 380 Thr Trp Gly Val Trp Pro Arg Pro Asp Trp Leu Leu Thr Ser Phe Trp 385 390 395 400 Gly Gly Asp Leu Arg Ala Ala Ser Asn Ile Ile Phe Ser Asn Gly Asn 405 410 415 Leu Asp Pro Trp Ala Gly Gly Gly Ile Arg Arg Asn Leu Ser Ala Ser 420 425 430 Val Ile Ala Val Thr Ile Gln Gly Gly Ala His His Leu Asp Leu Arg 435 440 445 Ala Ser His Pro Glu Asp Pro Ala Ser Val Val Glu Ala Arg Lys Leu 450 455 460 Glu Ala Thr Ile Ile Gly Glu Trp Val Lys Ala Ala Arg Arg Glu Gln 465 470 475 480 Gln Pro Ala Leu Arg Gly Gly Pro Arg Leu Ser Leu 485 490 3 1198 PRT Homo sapiens 3 Met Val Ala Ala Ala Ala Ala Thr Glu Ala Arg Leu Arg Arg Arg Thr 1 5 10 15 Ala Ala Thr Ala Ala Leu Ala Gly Arg Ser Gly Gly Pro His Cys Val 20 25 30 Asn Gly Gly Arg Cys Asn Pro Gly Thr Gly Gln Cys Val Cys Pro Ala 35 40 45 Gly Trp Val Gly Glu Gln Cys Gln His Cys Gly Gly Arg Phe Arg Leu 50 55 60 Thr Gly Ser Ser Gly Phe Val Thr Asp Gly Pro Gly Asn Tyr Lys Tyr 65 70 75 80 Lys Thr Lys Cys Thr Trp Leu Ile Glu Gly Gln Pro Asn Arg Ile Met 85 90 95 Arg Leu Arg Phe Asn His Phe Ala Thr Glu Cys Ser Trp Asp His Leu 100 105 110 Tyr Val Tyr Asp Gly Asp Ser Ile Tyr Ala Pro Leu Val Ala Ala Phe 115 120 125 Ser Gly Leu Ile Val Pro Glu Arg Asp Gly Asn Glu Thr Val Pro Glu 130 135 140 Val Val Ala Thr Ser Gly Tyr Ala Leu Leu His Phe Phe Ser Asp Ala 145 150 155 160 Ala Tyr Asn Leu Thr Gly Phe Asn Ile Thr Tyr Ser Phe Asp Met Cys 165 170 175 Pro Asn Asn Cys Ser Gly Arg Gly Glu Cys Lys Ile Ser Asn Ser Ser 180 185 190 Asp Thr Val Glu Cys Glu Cys Ser Glu Asn Trp Lys Gly Glu Ala Cys 195 200 205 Asp Ile Pro His Cys Thr Asp Asn Cys Gly Phe Pro His Arg Gly Ile 210 215 220 Cys Asn Ser Ser Asp Val Arg Gly Cys Ser Cys Phe Ser Asp Trp Gln 225 230 235 240 Gly Pro Gly Cys Ser Val Pro Val Pro Ala Asn Gln Ser Phe Trp Thr 245 250 255 Arg Glu Glu Tyr Ser Asn Leu Lys Leu Pro Arg Ala Ser His Lys Ala 260 265 270 Val Val Asn Gly Asn Ile Met Trp Val Val Gly Gly Tyr Met Phe Asn 275 280 285 His Ser Asp Tyr Asn Met Val Leu Ala Tyr Asp Leu Ala Ser Arg Glu 290 295 300 Trp Leu Pro Leu Asn Arg Ser Val Asn Asn Val Val Val Arg Tyr Gly 305 310 315 320 His Ser Leu Ala Leu Tyr Lys Asp Lys Ile Tyr Met Tyr Gly Gly Lys 325 330 335 Ile Asp Ser Thr Gly Asn Val Thr Asn Glu Leu Arg Val Phe His Ile 340 345 350 His Asn Glu Ser Trp Val Leu Leu Thr Pro Lys Ala Lys Glu Gln Tyr 355 360 365 Ala Val Val Gly His Ser Ala His Ile Val Thr Leu Lys Asn Gly Arg 370 375 380 Val Val Met Leu Val Ile Phe Gly His Cys Pro Leu Tyr Gly Tyr Ile 385 390 395 400 Ser Asn Val Gln Glu Tyr Asp Leu Asp Lys Asn Thr Trp Ser Ile Leu 405 410 415 His Thr Gln Gly Ala Leu Val Gln Gly Gly Tyr Gly His Ser Ser Val 420 425 430 Tyr Asp His Arg Thr Arg Ala Leu Tyr Val His Gly Gly Tyr Lys Ala 435 440 445 Phe Ser Ala Asn Lys Tyr Arg Leu Ala Asp Asp Leu Tyr Arg Tyr Asp 450 455 460 Val Asp Thr Gln Met Trp Thr Ile Leu Lys Asp Ser Arg Phe Phe Arg 465 470 475 480 Tyr Leu His Thr Ala Val Ile Val Ser Gly Thr Met Leu Val Phe Gly 485 490 495 Gly Asn Thr His Asn Asp Thr Ser Met Ser His Gly Ala Lys Cys Phe 500 505 510 Ser Ser Asp Phe Met Ala Tyr Asp Ile Ala Cys Asp Arg Trp Ser Val 515 520 525 Leu Pro Arg Pro Asp Ser Thr Met Met Ser Thr Asp Leu Ala Ile Pro 530 535 540 Ala Val Leu His Asn Ser Thr Met Tyr Val Phe Gly Gly Phe Asn Ser 545 550 555 560 Leu Leu Leu Ser Asp Ile Leu Val Phe Thr Ser Glu Gln Cys Asp Ala 565 570 575 His Arg Ser Glu Ala Ala Cys Leu Ala Ala Gly Pro Gly Ile Arg Cys 580 585 590 Val Trp Asn Thr Gly Ser Ser Gln Cys Ile Ser Trp Ala Leu Ala Thr 595 600 605 Asp Glu Gln Glu Glu Lys Leu Lys Ser Glu Cys Phe Ser Lys Arg Thr 610 615 620 Leu Asp His Asp Arg Cys Asp Gln His Thr Asp Cys Tyr Ser Cys Thr 625 630 635 640 Ala Asn Thr Asn Asp Cys His Trp Cys Asn Asp His Cys Val Pro Arg 645 650 655 Asn His Ser Cys Ser Glu Gly Gln Ile Ser Ile Phe Arg Tyr Glu Asn 660 665 670 Cys Pro Lys Asp Asn Pro Met Tyr Tyr Cys Asn Lys Lys Thr Ser Cys 675 680 685 Arg Ser Cys Ala Leu Asp Gln Asn Cys Gln Trp Glu Pro Arg Asn Gln 690 695 700 Glu Cys Ile Ala Leu Pro Glu Asn Ile Cys Gly Ile Gly Trp His Leu 705 710 715 720 Val Gly Asn Ser Cys Leu Lys Ile Thr Thr Ala Lys Glu Asn Tyr Asp 725 730 735 Asn Ala Lys Leu Phe Cys Arg Asn His Asn Ala Leu Leu Ala Ser Leu 740 745 750 Thr Thr Gln Lys Lys Val Glu Phe Val Leu Lys Gln Leu Arg Ile Met 755 760 765 Gln Ser Ser Gln Ser Met Ser Lys Leu Thr Leu Thr Pro Trp Val Gly 770 775 780 Leu Arg Lys Ile Asn Val Ser Tyr Trp Cys Trp Glu Asp Met Ser Pro 785 790 795 800 Phe Thr Asn Ser Leu Leu Gln Trp Met Pro Ser Glu Pro Ser Asp Ala 805 810 815 Gly Phe Cys Gly Ile Leu Ser Glu Pro Ser Thr Arg Gly Leu Lys Ala 820 825 830 Ala Thr Cys Ile Asn Pro Leu Asn Gly Ser Val Cys Glu Arg Pro Ala 835 840 845 Asn His Ser Ala Lys Gln Cys Arg Thr Pro Cys Ala Leu Arg Thr Ala 850 855 860 Cys Gly Asp Cys Thr Ser Gly Ser Ser Glu Cys Met Trp Cys Ser Asn 865 870 875 880 Met Lys Gln Cys Val Asp Ser Asn Ala Tyr Val Ala Ser Phe Pro Phe 885 890 895 Gly Gln Cys Met Glu Trp Tyr Thr Met Ser Thr Cys Pro Pro Glu Asn 900 905 910 Cys Ser Gly Tyr Cys Thr Cys Ser His Cys Leu Glu Gln Pro Gly Cys 915 920 925 Gly Trp Cys Thr Asp Pro Ser Asn Thr Gly Lys Gly Lys Cys Ile Glu 930 935 940 Gly Ser Tyr Lys Gly Pro Val Lys Met Pro Ser Gln Ala Pro Thr Gly 945 950 955 960 Asn Phe Tyr Pro Gln Pro Leu Leu Asn Ser Ser Met Cys Leu Glu Asp 965 970 975 Ser Arg Tyr Asn Trp Ser Phe Ile His Cys Pro Ala Cys Gln Cys Asn 980 985 990 Gly His Ser Lys Cys Ile Asn Gln Ser Ile Cys Glu Lys Cys Glu Asn 995 1000 1005 Leu Thr Thr Gly Lys His Cys Glu Thr Cys Ile Ser Gly Phe Tyr 1010 1015 1020 Gly Asp Pro Thr Asn Gly Gly Lys Cys Gln Pro Cys Lys Cys Asn 1025 1030 1035 Gly His Ala Ser Leu Cys Asn Thr Asn Thr Gly Lys Cys Phe Cys 1040 1045 1050 Thr Thr Lys Gly Val Lys Gly Asp Glu Cys Gln Leu Cys Glu Val 1055 1060 1065 Glu Asn Arg Tyr Gln Gly Asn Pro Leu Arg Gly Thr Cys Tyr Tyr 1070 1075 1080 Thr Leu Leu Ile Asp Tyr Gln Phe Thr Phe Ser Leu Ser Gln Glu 1085 1090 1095 Asp Asp Arg Tyr Tyr Thr Ala Ile Asn Phe Val Ala Thr Pro Asp 1100 1105 1110 Glu Gln Asn Arg Asp Leu Asp Met Phe Ile Asn Ala Ser Lys Asn 1115 1120 1125 Phe Asn Leu Asn Ile Thr Trp Ala Ala Ser Phe Ser Ala Gly Thr 1130 1135 1140 Gln Ala Gly Glu Glu Met Pro Val Val Ser Lys Thr Asn Ile Lys 1145 1150 1155 Glu Tyr Lys Asp Ser Phe Ser Asn Glu Lys Phe Asp Phe Arg Asn 1160 1165 1170 His Pro Asn Ile Thr Phe Phe Val Tyr Val Ser Asn Phe Thr Trp 1175 1180 1185 Pro Ile Lys Ile Gln Val Gln Thr Glu Gln 1190 1195 4 310 PRT Homo sapiens 4 Phe Glu Gly Thr Lys Asp Ser Pro Leu Glu His His Leu Tyr Val Val 1 5 10 15 Ser Tyr Val Asn Pro Gly Glu Val Thr Arg Leu Thr Asp Arg Gly Tyr 20 25 30 Ser His Ser Cys Cys Ile Ser Gln His Cys Asp Phe Phe Ile Ser Lys 35 40 45 Tyr Ser Asn Gln Lys Asn Pro His Cys Val Ser Leu Tyr Lys Leu Ser 50 55 60 Ser Pro Glu Asp Asp Pro Thr Cys Lys Thr Lys Glu Phe Trp Ala Thr 65 70 75 80 Ile Leu Asp Ser Ala Gly Pro Leu Pro Asp Tyr Thr Pro Pro Glu Ile 85 90 95 Phe Ser Phe Glu Ser Thr Thr Gly Phe Thr Leu Tyr Gly Met Leu Tyr 100 105 110 Lys Pro His Asp Leu Gln Pro Gly Lys Lys Tyr Pro Thr Val Leu Phe 115 120 125 Ile Tyr Gly Gly Pro Gln Gly Gln Ile Glu Ile Asp Asp Gln Val Glu 130 135 140 Gly Leu Gln Tyr Leu Ala Ser Arg Tyr Asp Phe Ile Asp Leu Asp Arg 145 150 155 160 Val Gly Ile His Gly Trp Ser Tyr Gly Gly Tyr Leu Ser Leu Met Ala 165 170 175 Leu Met Gln Arg Ser Asp Ile Phe Arg Val Ala Ile Ala Gly Ala Pro 180 185 190 Val Thr Leu Trp Ile Phe Tyr Asp Thr Gly Tyr Thr Glu Arg Tyr Met 195 200 205 Gly His Pro Asp Gln Asn Glu Gln Gly Tyr Tyr Leu Gly Ser Val Ala 210 215 220 Met Gln Ala Glu Lys Phe Pro Ser Glu Pro Asn Arg Leu Leu Leu Leu 225 230 235 240 His Gly Phe Leu Asp Glu Asn Val His Phe Ala His Thr Ser Ile Leu 245 250 255 Leu Ser Phe Leu Val Arg Ala Gly Lys Pro Tyr Asp Leu Gln Ile Tyr 260 265 270 Pro Gln Glu Arg His Ser Ile Arg Val Pro Glu Ser Gly Glu His Tyr 275 280 285 Glu Leu His Leu Leu His Tyr Leu Gln Glu Asn Leu Gly Ser Arg Ile 290 295 300 Ala Ala Leu Lys Val Ile 305 310 

What is claimed is:
 1. A method of treating a patient for mucosal inflammation associated with rhinitis, sinusitis or both, comprising: intranasally administering to said patient a therapeutically effective amount of a peptidase that cleaves at Xaa-Pro sequences.
 2. The method of claim 1, wherein said peptidase is an exopeptidase selected from the group consisting of: dipeptidyl peptidase IV; quiescent cell proline dipeptidase; dipeptidyl peptidase 8; and attractin.
 3. The method of claim 2, wherein said exopeptidase is dipeptidyl peptidase IV.
 4. The method of claim 2, wherein said exopeptidase is dipeptidyl peptidase
 8. 5. The method of claim 2, wherein said exopeptidase is quiescent cell proline dipeptidase.
 6. The method of claim 2, wherein said exopeptidase is attractin.
 7. The method of claim 2, wherein said exopeptidase is administered at a dose of between 1 μg and 1 mg.
 8. The method of any one of claims 1-7, wherein said rhinitis, sinusitis or both, is the result of allergies or asthma.
 9. A therapeutic package comprising a device for intranasally delivering drug to a patient, preloaded with a solution or powder comprising a peptidase that cleaves at Xaa-Pro sequences.
 10. The therapeutic package of claim 9, wherein said peptidase is an exopeptidase selected from the group consisting of: dipeptidyl peptidase IV; quiescent cell proline dipeptidase; dipeptidyl peptidase 8; and attractin.
 11. The therapeutic package of claim 9, wherein said exopeptidase is dipeptidyl peptidase IV.
 12. The therapeutic package of claim 9, wherein said exopeptidase is quiescent cell proline dipeptidase.
 13. The therapeutic package of claim 10, wherein said exopeptidase is dipeptidyl peptidase
 8. 14. The therapeutic package of claim 9, wherein said exopeptidase is attractin.
 15. The therapeutic package of claim 10, wherein said exopeptidase is present in a solution at a concentration of between 1 μg/ml and 10 mg/ml.
 16. A method of treating a patient for mucosal inflammation associated with rhinitis, sinusitis or both, comprising: intranasally administering to said patient a therapeutically effective amount of an agent that inhibits the binding of SP to the NK1 receptor. 